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Sökning: WFRF:(Kreij Karl 1975 )

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1.
  • Kreij, Karl, 1975- (författare)
  • Bioprocess monitoring of mammalian cell cultures using modern sensors
  • 2003
  • Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
    • The need for better monitoring and control of mammalian cell cultures and the quality and safety control of products obtained from these processes is still high. In an attempt to enable better monitoring of the process and the glycosylation pattern of typical mammalian cell cultures two modem sensor methods were used on a chinese hamster ovary (CHO) cell culture producing the glycoprotein macrophage colony stimulating factor (M-CSF). An electronic nose was connected to the reactor and measurements were made online. Surface plasmon resonance (SPR) measurement were performed on cell culture supernatant samples.The electronic nose was used to study microbial and fungal infections in a cell culture. Principal component analysis and cluster analysis were used to evaluate the multivariate data obtained from the electronic nose. The results indicate that it is possible to use the electronic nose to detect contaminations in cell cultures.A novel method that combines the strong affinity of an antibody with the weak affinity of lectins was used in SPR measurements of the crude cell culture supernatant. It is suggested that this method could be used to monitor the glycosylation pattern of recombinant glycoproteins on-line.
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2.
  • Kreij, Karl, 1975-, et al. (författare)
  • On-line detection of microbial contaminations in animal cell reactor cultures using an electronic nose device
  • 2005
  • Ingår i: Cytotechnology (Dordrecht). - : Springer Science and Business Media LLC. - 0920-9069 .- 1573-0778. ; 48:1-3, s. 41-58
  • Tidskriftsartikel (refereegranskat)abstract
    • An electronic nose (EN) device was used to detect microbial and viral contaminations in a variety of animal cell culture systems. The emission of volatile components from the cultures accumulated in the bioreactor headspace, was sampled and subsequently analysed by the EN device. The EN, which was equipped with an array of 17 chemical gas sensors of varying selectivity towards the sampled volatile molecules, generated response patterns of up to 85 computed signals. Each 15 or 20 min a new gas sample was taken generating a new response pattern. A software evaluation tool visualised the data mainly by using principal component analysis. The EN was first used to detect microbial contaminations in a Chinese hamster ovary (CHO) cell line producing a recombinant human macrophage colony stimulating factor (rhM-CSF). The CHO cell culture was contaminated by Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida utilis which all were detected. The response patterns from the CHO cell culture were compared with monoculture references of the microorganisms. Second, contaminations were studied in an Sf-9 insect cell culture producing another recombinant protein (VP2 protein). Contaminants were detected from E. coli, a filamentous fungus and a baculovirus. Third, contamination of a human cell line, HEK-293, infected with E. coli exhibited comparable results. Fourth, bacterial contaminations could also be detected in cultures of a MLV vector producer cell line. Based on the overall experiences in this study it is concluded that the EN method has in a number of cases the potential to be developed into a useful on-line contamination alarm in order to support safety and economical operation for industrial cultivation. © Springer 2005.
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